• The Korean journal of helicobacter and upper gastrointestinal research
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• v.18 no.3
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• pp.157-161
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• 2018

Liquid biopsy, the analysis of circulating biomarkers from peripheral blood, such as circulating tumor cells (CTCs) and circulating tumor DNA, and exosomes, offers a less invasive, new source of cancer-derived materials that may reflect the status of the disease better and thereby contribute to personalized treatment. Recent advances in microfluidics and molecular analysis technologies have resulted in greatly improved CTC enumeration and detection. In this article, we review commercially available technologies used to isolate CTCs from peripheral blood, including immunoaffinity and label-free, physical property-based isolation methods. Although enormous technological progress has been made, especially within the last decade, only a few CTC detection methods have been approved for routine clinical use. Here, we provide an overview of the current CTC isolation methods and examples of their potential application for early diagnosis, prognosis, treatment monitoring, and prediction of resistance to cancer therapy. Furthermore, the challenges that remain to be addressed before such tools are implemented for routine use in clinical settings are discussed.

• Genomics & Informatics
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• v.17 no.4
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• pp.42.1-42.6
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• 2019

Robust identification of genetic alterations is important for the diagnosis and subsequent treatment of tumors. Screening for genetic alterations using tumor tissue samples may lead to biased interpretations because of the heterogeneous nature of the tumor mass. Liquid biopsy has been suggested as an attractive tool for the non-invasive follow-up of cancer treatment outcomes. In this study, we aimed to verify whether the mutations identified in primary tumor tissue samples could be consistently detected in plasma cell-free DNA (cfDNA) by digital polymerase chain reaction (dPCR). We first examined the genetic alteration profiles of three colorectal cancer (CRC) tissue samples by targeted next-generation sequencing (NGS) and identified 11 non-silent amino acid changes across six cancer-related genes (APC, KRAS, TP53, TERT, ARIDIA, and BRCA1). All three samples had KRAS mutations (G12V, G12C, and G13D), which were well-known driver events. Therefore, we examined the KRAS mutations by dPCR. When we examined the three KRAS mutations by dPCR using tumor tissue samples, all of them were consistently detected and the variant allele frequencies (VAFs) of the mutations were almost identical between targeted NGS and dPCR. When we examined the KRAS mutations using the plasma cfDNA of the three CRC patients by dPCR, all three mutations were consistently identified. However, the VAFs were lower (range, 0.166% to 2.638%) than those obtained using the CRC tissue samples. In conclusion, we confirmed that the KRAS mutations identified from CRC tumor tissue samples were consistently detected in the plasma cfDNA of the three CRC patients by dPCR.

• BMB Reports
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• v.56 no.3
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• pp.190-195
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• 2023

We propose a novel blood biomarker detection method that uses miRNA super-resolution imaging to enable the early diagnosis of Alzheimer's disease (AD). Here, we report a single-molecule detection method for visualizing disease-specific miRNA in tissue from an AD mice model, and peripheral blood mononuclear cells (PBMCs) from AD patients. Using optimized Magnified Analysis of Proteome (MAPs), we confirmed that five miRNAs contribute to neurodegenerative disease in the brain hippocampi of 5XFAD and wild-type mice. We also assessed PBMCs isolated from the whole blood of AD patients and a healthy control group, and subsequently analyzed those samples using miRNA super-resolution imaging. We detected more miR-200a-3p expression in the cornu ammonis 1 and dentate gyrus regions of 3 month-old 5XFAD mice than in wild-type mice. Additionally, miRNA super-resolution imaging of blood provides AD diagnosis platform for studying miRNA regulation inside cells at the single molecule level. Our results present a potential liquid biopsy method that could improve the diagnosis of early stage AD and other diseases.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.9
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• pp.4567-4570
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• 2012

Objectives: The purposes of this study were to determine the prevalence and predictive value to detect significant neoplasia and invasive lesions, and to evaluate the correlation between clinical and histopathology of women with squamous cell carcinoma (SCCA) on Siriraj liquid-based cervical cytology (Siriraj-LBC). Methods: The computerized database of women who underwent Siriraj-LBC at Siriraj Hospital, Mahidol University from January 2007 to December 2010 were retrieved. The hospital records of women with SCCA cytology were reviewed. Results: The prevalence of SCCA cytology was 0.07%. A total of 86 women, mean age was 58.1 years. Sixty-one women (70.9%) were post-menopausal. Overall significant pathology and invasive gynecologic cancer were detected in 84 women (97.7%) and 71 women (82.5%), respectively. The positive predictive values for detection of significant neoplasia and invasive lesion were 97.7% and 82.6%, respectively. The cervical cancer was diagnosed in 69 women and among these 58 women were SCCA. Thirteen women (15.1%) had cervical intraepithelial neoplasia (CIN) 3 and two women (2.3%) had cervicitis. The sensitivity and specificity of colposcopy for cervical cancer detection in SCCA cytology were 83.3% and 75%, respectively. Median follow up period was 17.6 months and 64 patients were alive without cytologic abnormality. Conclusions: The final histopathology of SCCA cytology in our populations demonstrated a wide variety, from cervicitis to invasive cancer and the most common diagnosis was invasive cervical cancer. Colposcopy with biopsy and/or endocervical curettage and loop electrosurgical excision procedure should be undertaken to achieve histologic diagnosis.

• The Korean Journal of Cytopathology
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• v.15 no.1
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• pp.33-39
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• 2004

We compared the diagnostic accuracy of liquid-based cervicovaginal cytology using $MonoPrep2^{TM}$ system (Monogen, Herndon, Virginia, USA), a manual system based on membrane filtration method, with conventional Pap smear. Study population included 92 patients visiting the gynecologic department under the suspicion of uterine cervical disease. In thirty of them, surgical biopsy was performed. $MonoPrep2^{TM}$ system provided well-preserved monolayer specimen with good nuclear morphology. However, about 19% of specimens were inadequate to interpret due to low cellularity. The detection rate of abnormal cells more than ASCUS (atypical squamous cells of unknown significance) was 23.9% and higher than 19.4 % of conventional Pap smear. Diagnostic concordance rate with conventional Pap smear was 81%, and severe discordance rate influencing on the management of patient was 7.6 %. Among these seven cases, $MonoPrep2^{TM}$ system was more diagnostic only in four. In comparison with histology, the sensitivity of diagnosis of $MonoPrep2^{TM}$ system was 78.9% and slightly higher than 73.5% of conventional Pap smear. However, the specificity was 81.1% and lower than 90.9% of Pap smear. In conclusion, $MonoPrep2^{TM}$ system provided diagnostic accuracies similar to the conventional Pap smear. The inexpertness of slide preparation and the low cellularity were considered to endow a limitation in more accurate evaluation.

• Biomedical Science Letters
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• v.15 no.1
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• pp.47-53
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• 2009

Urinary bladder cancer is diagnosed through urine cytology and cytoscopy with biopsy. An atypical urothelial cast is often found by voided urine cytology in a papillary urothelial cell carcinoma. The objective of this study is to demonstrate the significance of the evaluation of urinary nuclear matrix protein (NMP22) level and Sure Path Liquid-based cytology (SP-LBC) as compared to the examination of atypical urothelial cast in voided urine sediment for monitoring bladder cancer. From October 2007 to January 2008, we observed 3240 patients who visited the emergency laboratory of urology of Soonchunhyang University, Cheonan Hospital. Both NMP22 measurement and SP-LBC were performed in 31 patients who were positive in an atypical urothelial cast test. In particular, 26 men and 5 women were found to be atypical urothelial cast-positive persons. The average age for both men and women is 61.8. NMP22 test is positive in 23 of 31 cases (74.2%) from patients with atypical urothelial cast, while the test is negative in 8 of 31 cases (25.8%). The percentages of negativity, atypicality, suspicious malignancy, and malignancy in SP-LBC are 25.8% (8/31), 58.1% (18/31), 9.7% (3/31), and 6.5% (2/31), respectively. The relation of NMP22 positivity with the malignant degree in LBC is significant (P<0.01). Two malignant patients resulting from SP-LBC show the same results in histological examination. Overall, the study suggests the usefulness of NMP22 measurement and LBC as well as the examination of atypical urothelial cast for the diagnosis of early bladder cancer.

• Tuberculosis and Respiratory Diseases
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• v.78 no.2
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• pp.64-71
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• 2015

Pulmonary tuberculosis (TB) persists as a great public health problem in Korea. Increases in the overall age of the population and the rise of drug-resistant TB have reinforced the need for rapid diagnostic improvements and new modalities to detect TB and drug-resistant TB, as well as to improve TB control. Standard guidelines and recent advances for diagnosing pulmonary TB are summarized in this article. An early and accurate diagnosis of pulmonary TB should be established using chest X-ray, sputum microscopy, culture in both liquid and solid media, and nucleic acid amplification. Chest computed tomography, histopathological examination of biopsy samples, and new molecular diagnostic tests can be used for earlier and improved diagnoses, especially in patients with smear-negative pulmonary TB or clinically-diagnosed TB and drug-resistant TB.

• The Korean Journal of Cytopathology
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• v.18 no.2
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• pp.119-125
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• 2007

Urine cytology is an important screening tool for urinary tract neoplasms. Liquid-based preparation methods, such as $ThinPrep^{(R)}$, have been introduced for non-gynecological samples. We aimed to assess the diagnostic accuracy of liquid-based preparations in urine cytology by comparing the results of the conventional Cytospin preparation method for the same samples. A total of 236 cases subject to urine cytology were enrolled in this study from January 2005 to December 2005. All cases were subjected to cystoscopy and if a malignancy was suspected, a biopsy was performed. Urine cytology slides were made using the $ThinPrep^{(R)}$ preparation method and the conventional Cytospin and/or direct smear method from the individual samples. The results of urine cytology were compared with the final cystoscopic or histological diagnoses. We analyzed the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of both cytology preparation methods. A total of 236 slides made using the liquid based method were satisfactory for slide quality, whereas 5 slides (2.1%) prepared by conventional methods were unsatisfactory because of air-drying, a thick smear, or a bloody or inflammatory background. The $ThinPrep^{(R)}$ method showed 53.1% sensitivity, 92.6% specificity, a 92,6% positive predictive value, a 94.1% negative predictive value and 85,6% accuracy, while the conventional method showed 51% sensitivity, 98.4% specificity, a 92.6% positive predictive value, a 98.4% negative predictive value and 88,6% accuracy. Although the diagnostic values were equivalent between the use of the two methods, the quality of the cytology slides and the time consumed during the microscopic examination for a diagnosis were superior for the $ThinPrep^{(R)}$ method than for the conventional method. In conclusion, our limited studies have shown that the use of the liquid based preparation method is beneficial to improve the quality of slides and reduce the duration for a microscopic examination, but did not show better sensitivity, accuracy and predictive values.

• Journal of Nutrition and Health
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• v.24 no.3
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• pp.199-205
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• 1991

Retinoic acid. the active metabolite of vitamin A. was detected in the human liver for the first time using a new method. A rapid and sensitive technique has been developed using gradient-elution. reverse-phase high performance liquid chromatography. This assay. with simultaneous multiwavelength detection at 294nm, 325nm and 450nm after saponifcation of liver samples. allows us seperation and quantitation of vitamin E, retinoic acid, total retinoids and various carotenoids in one small sample. The proportion of retinoic acid to total retinoids in human liver appears to be quite $consistent(2.4\pm0.2%$ ). With low vitamin A storage in liver, detection at another wavelenth 354nm would increase the sensitivity for retinoic acid of small quantity This method of analysis could be used for other tissues like red blood cells, plasma or serum, also. Hepatic retinoic acid level with total retinoids and carotenoids would serve a better indicator of functional vitamin A nutriture especially for those with disease requiring needle biopsy of liver.

• The Korean Journal of Cytopathology
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• v.19 no.2
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• pp.119-125
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• 2008

This study was performed to compare the efficacy between a DNA chip method and a Hybrid-Capture II assay (HC-II) for detecting human papillomavirus in patients with intraepithelial lesions of the uterine cervix. From May, 2005, to June, 2006, 192 patients with abnormal colposcopic findings received cervical cytology, HC-II and HPV DNA chip tests, and colposcopic biopsy or conization. We compared the results of HC-II and HPV DNA chip in conjunction with liquid based cervical cytology (LBCC) and confirmed the results of biopsy or conization. The sensitivity of the HPV DNA chip test was higher than HC-II or LBCC. The HPV DNA chip in conjunction with LBCC showed higher sensitivity than any single method and higher sensitivity than HC-II with LBCC. We confirmed that the HPV DNA chip test was more sensitive for detecting HPV in cervical lesions than HC-II, and that it would provide more useful clinical information about HPV type and its multiple infections.